Investigating the biotechnological potential of thermophiles using integrated culture-dependent and metagenomic approaches from Tapovan hot springs for industrial thermozymes

Hot springs are prolific reservoirs for bioprospecting novel biomolecules with industrial relevance. The present study reports the first integrated culture-dependent and culture enriched and sequencing based assessment of microbial diversity and metabolic potential at the Tapovan hot spring (Uttarakhand, India). Fifteen genera belonging to five major phyla were identified from soil and water samples using both culture-dependent culture-enriched and culture-independent (16S amplicon sequencing) approaches. Based on 16S rRNA gene sequencing of culturable isolates, Paenibacillus phoenicis , Paenibacillus naphthalenovorans , Paenibacillus faecis , and Aeribacillus pallidus were preliminarily identified; their presence at Tapovan extends the known geographical and ecological range of these species. Additional genome-based research will be necessary to confirm their taxonomic identity and ecological importance. Qualitative and quantitative enzyme screening revealed that isolates produced key industrial enzymes including amylase, cellulase, xylanase, mannanase, and pectinase. Importantly, most characterised strains demonstrated the ability to produce multiple enzymes simultaneously, reflecting broad metabolic versatility. These findings demonstrate the potential of Tapovan hot spring as a source of thermostable, multi-functional enzymes, and indicate the need for further genome level characterisation of its thermophilic community for biotechnological exploitation.